LOCUS       sequence-367916-        9423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLT3 D835Y activating tyrosine kinase domain mutant in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9423)
  TITLE     Mammalian expression plasmids for FLT3 wildtype and common AML 
            mutant FLT3 proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: FLT3-D835Y-MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..9423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4415..4982
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4432..4984
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4576..4593)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4803..4822
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4996..4998,4999..5001,5002..5715)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5041..5062)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5302..5321)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5649..5670
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5817..6331
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6493..6509
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6517..6523,6524..6541,6542..6547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6562..6583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6700..6717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6871..7459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6951..6970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7630..8421,8422..8490))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8253..8272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8491..8595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8663..8681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8719..8741)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8841..8860
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9054..9076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9204..9223
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(9423,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctatgc cggcgttggc gcgcgacggc
     1441 ggccagctgc cgctgctcgt tgttttttct gcaatgatat ttgggactat tacaaatcaa
     1501 gatctgcctg tgatcaagtg tgttttaatc aatcataaga acaatgattc atcagtgggg
     1561 aagtcatcat catatcccat ggtatcagaa tccccggaag acctcgggtg tgcgttgaga
     1621 ccccagagct cagggacagt gtacgaagct gccgctgtgg aagtggatgt atctgcttcc
     1681 atcacactgc aagtgctggt cgatgcccca gggaacattt cctgtctctg ggtctttaag
     1741 cacagctccc tgaattgcca gccacatttt gatttacaaa acagaggagt tgtttccatg
     1801 gtcattttga aaatgacaga aacccaagct ggagaatacc tactttttat tcagagtgaa
     1861 gctaccaatt acacaatatt gtttacagtg agtataagaa ataccctgct ttacacatta
     1921 agaagacctt actttagaaa aatggaaaac caggacgccc tggtctgcat atctgagagc
     1981 gttccagagc cgatcgtgga atgggtgctt tgcgattcac agggggaaag ctgtaaagaa
     2041 gaaagtccag ctgttgttaa aaaggaggaa aaagtgcttc atgaattatt tgggatggac
     2101 ataaggtgct gtgccagaaa tgaactgggc agggaatgca ccaggctgtt cacaatagat
     2161 ctaaatcaaa ctcctcagac cacattgcca caattatttc ttaaagtagg ggaaccctta
     2221 tggataaggt gcaaagctgt tcatgtgaac catggattcg ggctcacctg ggaattagaa
     2281 aacaaagcac tcgaggaggg caactacttt gagatgagta cctattcaac aaacagaact
     2341 atgatacgga ttctgtttgc ttttgtatca tcagtggcaa gaaacgacac cggatactac
     2401 acttgttcct cttcaaagca tcccagtcaa tcagctttgg ttaccatcgt agaaaaggga
     2461 tttataaatg ctaccaattc aagtgaagat tatgaaattg accaatatga agagttttgt
     2521 ttttctgtca ggtttaaagc ctacccacaa atcagatgta cgtggacctt ctctcgaaaa
     2581 tcatttcctt gtgagcaaaa gggtcttgat aacggataca gcatatccaa gttttgcaat
     2641 cataagcacc agccaggaga atatatattc catgcagaaa atgatgatgc ccaatttacc
     2701 aaaatgttca cgctgaatat aagaaggaaa cctcaagtgc tcgcagaagc atcggcaagt
     2761 caggcgtcct gtttctcgga tggataccca ttaccatctt ggacctggaa gaagtgttca
     2821 gacaagtctc ccaactgcac agaagagatc acagaaggag tctggaatag aaaggctaac
     2881 agaaaagtgt ttggacagtg ggtgtcgagc agtactctaa acatgagtga agccataaaa
     2941 gggttcctgg tcaagtgctg tgcgtacaat tcccttggca catcttgtga gacgatcctt
     3001 ttaaactctc caggcccctt ccctttcatc caagacaaca tctcattcta tgcaacaatt
     3061 ggtgtttgtc tcctcttcat tgtcgtttta accctgctaa tttgtcacaa gtacaaaaag
     3121 caatttaggt atgaaagcca gctacagatg gtacaggtga ccggctcctc agataatgag
     3181 tacttctacg ttgatttcag agaatatgaa tatgatctca aatgggagtt tccaagagaa
     3241 aatttagagt ttgggaaggt actaggatca ggtgcttttg gaaaagtgat gaacgcaaca
     3301 gcttatggaa ttagcaaaac aggagtctca atccaggttg ccgtcaaaat gctgaaagaa
     3361 aaagcagaca gctctgaaag agaggcactc atgtcagaac tcaagatgat gacccagctg
     3421 ggaagccacg agaatattgt gaacctgctg ggggcgtgca cactgtcagg accaatttac
     3481 ttgatttttg aatactgttg ctatggtgat cttctcaact atctaagaag taaaagagaa
     3541 aaatttcaca ggacttggac agagattttc aaggaacaca atttcagttt ttaccccact
     3601 ttccaatcac atccaaattc cagcatgcct ggttcaagag aagttcagat acacccggac
     3661 tcggatcaaa tctcagggct tcatgggaat tcatttcact ctgaagatga aattgaatat
     3721 gaaaaccaaa aaaggctgga agaagaggag gacttgaatg tgcttacatt tgaagatctt
     3781 ctttgctttg catatcaagt tgccaaagga atggaatttc tggaatttaa gtcgtgtgtt
     3841 cacagagacc tggccgccag gaacgtgctt gtcacccacg ggaaagtggt gaagatatgt
     3901 gactttggat tggctcgata tatcatgagt gattccaact atgttgtcag gggcaatgcc
     3961 cgtctgcctg taaaatggat ggcccccgaa agcctgtttg aaggcatcta caccattaag
     4021 agtgatgtct ggtcatatgg aatattactg tgggaaatct tctcacttgg tgtgaatcct
     4081 taccctggca ttccggttga tgctaacttc tacaaactga ttcaaaatgg atttaaaatg
     4141 gatcagccat tttatgctac agaagaaata tacattataa tgcaatcctg ctgggctttt
     4201 gactcaagga aacggccatc cttccctaat ttgacttcgt ttttaggatg tcagctggca
     4261 gatgcagaag aagcgatgta tcagaatgtg gatggccgtg tttcggaatg tcctcacacc
     4321 taccaaaaca ggcgaccttt cagcagagag atggatttgg ggctactctc tccgcaggct
     4381 caggtcgaag attcgtagct cgaggttaac gaattccgcc ccccccccct aacgttactg
     4441 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     4501 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     4561 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     4621 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     4681 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     4741 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     4801 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     4861 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     4921 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     4981 aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     5041 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag
     5101 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     5161 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     5221 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     5281 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     5341 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     5401 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     5461 gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc
     5521 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     5581 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     5641 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     5701 gagctgtaca agtaatgaat taattaagaa ttatcaagct tatcgatacc gtcgacctgc
     5761 agccaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     5821 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     5881 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     5941 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     6001 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     6061 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     6121 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     6181 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     6241 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     6301 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     6361 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     6421 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     6481 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8701 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8761 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8821 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     8881 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     8941 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9001 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9061 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     9121 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     9181 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     9241 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     9301 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     9361 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     9421 gga
//
